淡紫紫孢菌的筛选、鉴定及产壳聚糖酶固体发酵条件优化
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“十三五”国家重点研发计划重点专项(2018YFD0400302); 中央高校基本科研业务费专项(2662019PY032)


Isolation and Identification of Purpureocillium lilacinum and Optimization of It’s Solid Fermentation Conditions for Producing Chitosanase
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    目的:从全国各地土样中分离纯化出一株产壳聚糖酶活性较高的真菌M7a,并优化其固体发酵产酶条件。方法:综合形态学观察和18S rDNA序列测定进行鉴定,采用单因素和响应面设计确定其最佳固体发酵产壳聚糖酶条件。结果:菌株M7a被鉴定为淡紫紫孢菌,其最佳固体发酵产壳聚糖酶条件为:10 g/L胶体壳聚糖+5 g/L葡萄糖,10 g/L蛋白胨,豆粕麸皮质量比7 ∶ 5,初始pH值为6.0,发酵温度33 ℃及发酵时间5.5 d。产壳聚糖酶优化后达到16.80 U/mL,是初始条件的5.1倍。SDS-PAGE和酶谱分析发现该菌株产胞外分子质量为40.0 ku的壳聚糖酶,且无同工酶。结论:本研究筛选鉴定出一株新颖且产壳聚糖酶活力较高的淡紫紫孢菌 M7a,优化了固体发酵条件,提高了产酶水平,为淡紫紫孢菌壳聚糖酶的分离纯化和应用提供了理论基础。

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    Objective: A fungal with high activity of chitosanase was isolated from soil samples and named as M7a. Then, optimize solid fermentation conditions of the strain M7a for chitosanase production. Method: The strain M7a was identified based on its morphological characters and 18S rDNA sequence. The single factor and response surface test were used to determine it’s optimum solid fermentation conditions for chitosanase production. Result: The strain M7a was identified as Purpureocillium lilacinum. The optimum solid fermentation conditions were confirmed as follows: 10 g/L colloidal chitosan + 5 g/L glucose, 10 g/L peptone, the mass ratio of soybean meal was 7 ∶ 5, the initial pH of the medium was 6.0, 33 ℃ for 5.5 d. The final chitosanase activity reached 16.80 U/mL, which was 5.1 folds higher than the beginning after optimization. SDS-PAGE and zymography analysis showed that the strain M7a secreted a 40 ku chitosanase without isozyme. Conclusion: This study screened and identified a novel strain of Purpureocillium lilacinum M7a with high chitosanase activity, and greatly improved the chitosanase production level of solid fermentation. The results can provide a theoretical basis for purification and application of the chitosanase from Purpureocillium lilacinum.

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张馨月;岳晓洁;李铮峥;刘倩;黄茜;马美湖;付星;.淡紫紫孢菌的筛选、鉴定及产壳聚糖酶固体发酵条件优化[J].中国食品学报,2020,20(4):160-169

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  • 在线发布日期: 2020-04-30
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